Method for refining protamine sulfate
A technology of protamine sulfate and protamine, applied in the field of biomedicine, can solve the problem of high fatality rate
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[0020] The protamine essence of salmon is crushed, extracted with water, filtered, and the filtrate is precipitated with ethanol to obtain the crude product of protamine sulfate, which is then subjected to the following operations:
[0021] (1) Protamine enzymatic hydrolysis process
[0022] Accurately weigh 50mg of protamine sulfate crude product, dissolve in 50ml CaCl 2 In the buffer solution, continue to stir and dissolve for about 1 hour until completely dissolved. Accurately weigh 5 mg of thermolysin, and add it into a fully dissolved protamine buffer to obtain a reaction solution with a protamine:thermolysin (w / w) ratio of 100:1. Filter and set aside.
[0023] (2) Separation and purification process
[0024] Separate and weigh Tris buffer salt and NaCl to prepare mobile phase A; 50mM Tris solution and mobile phase B:, 50mM Tris / 2MNaCl solution. The pH was adjusted to 7.5 with HCl. Install the heparin affinity chromatographic column on the fast protein chromatography...
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